LOCUS       sequence-311095-        4900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast tagging vector to create gene fusion with yeast codon 
            optimized mTFP1 under KanMX selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4900)
  TITLE     Stelling Lab Plasmids
REFERENCE   2  (bases 1 to 4900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: FRP265
FEATURES             Location/Qualifiers
     source          1..4900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..74)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     174..193
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        274..292
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     274..291
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      1105..1292
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1160..1184)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1367..2723
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1367..1710
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1711..2520
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1778..1797)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2526..2723
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2827..2846)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2828..2846)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2933..2950)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3104..3692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3184..3203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3863..4654,4655..4723))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4486..4505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4724..4828)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(4896..4900,1..14)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
       61 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
      121 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
      181 agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa
      241 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaacgcggcc
      301 gccagctgaa gcttcgtacg ctgcaggtcg acggatcggt gacggtgctg gtttaattaa
      361 caccggtatg gtttctaaag gtgaagaaac tactatgggt gttattaaac cagatatgaa
      421 gattaagttg aagatggaag gtaatgttaa tggtcatgct tttgttattg aaggtgaagg
      481 tgaaggtaaa ccatatgatg gtacaaacac tattaacttg gaagttaaag aaggtgctcc
      541 attgccattt tcttacgata ttttgactac tgcttttgct tatggtaata gagcttttac
      601 taagtaccca gatgatattc caaattactt caagcaatct tttccagaag gttattcttg
      661 ggaaagaact atgacttttg aagataaggg tattgttaag gttaagtctg atatttctat
      721 ggaagaagat tctttcattt acgaaattca tttgaagggt gaaaattttc caccaaatgg
      781 tccagttatg caaaaaaaga ctactggttg ggatgcttct actgaaagga tgtacgtcag
      841 agatggtgtt ttgaaaggtg atgttaagca taaattgttg ttggaaggtg gtggtcatca
      901 tagagttgat ttcaagacaa tttacagagc taaaaaggct gttaaattgc cagattacca
      961 ttttgttgat catagaattg aaattttgaa ccatgataag gattacaaca aggttactgt
     1021 ttatgaatct gctgttgcta gaaattctac tgatggtatg gatgaattgt ataaagccgg
     1081 ctagggcgcg ccacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     1141 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     1201 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat
     1261 gaggtcgctc ttattgacca cacctctacc ggcagatccg ctagggataa cagggtaata
     1321 tagatctgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca
     1381 gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc
     1441 gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg
     1501 cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc
     1561 cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta
     1621 ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag
     1681 ttctcacatc acatccgaac ataaacaacc atgggtaagg aaaagactca cgtttcgagg
     1741 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1801 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
     1861 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     1921 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1981 gatgcatggt tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa
     2041 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     2101 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     2161 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     2221 tggcctgttg aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca
     2281 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     2341 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2401 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2461 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     2521 tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt
     2581 atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg
     2641 acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta
     2701 tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa
     2761 cgagctcgaa ttcatcgatg atatcagatc cactagtggc ctatgcggcc gcggatctgc
     2821 cggtctccct atagtgagtc gtattaattt cgataagcca ggttaacctg cattaatgaa
     2881 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2941 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3001 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3061 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3121 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3181 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3241 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3301 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3361 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3421 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3481 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3541 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3601 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3661 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3721 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3781 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3841 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3901 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3961 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4021 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4081 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4141 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4201 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4261 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4321 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4381 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4441 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4501 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4561 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4621 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4681 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4741 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4801 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     4861 aagaaaccat tattatcatg acattaacct ataaaaatag
//
